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Open AccessJournal ArticleDOI

Dynamics in the plasma membrane: how to combine fluidity and order

TLDR
The basic concepts of Brownian diffusion and lipid domain formation in model membranes are summarized and the development of ideas and tools in this field are tracked, outlining key results obtained on the dynamic processes at work in membrane structure and assembly.
Abstract
Cell membranes are fascinating supramolecular aggregates that not only form a barrier between compartments but also harbor many chemical reactions essential to the existence and functioning of a cell. Here, it is proposed to review the molecular dynamics and mosaic organization of the plasma membrane, which are thought to have important functional implications. We will first summarize the basic concepts of Brownian diffusion and lipid domain formation in model membranes and then track the development of ideas and tools in this field, outlining key results obtained on the dynamic processes at work in membrane structure and assembly. We will focus in particular on findings made using fluorescent labeling and imaging procedures to record these dynamic processes. We will also discuss a few examples showing the impact of lateral diffusion on cell signal transduction, and outline some future methodological challenges which must be met before we can answer some of the questions arising in this field of research.

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Journal ArticleDOI

Lipid rafts: at a crossroad between cell biology and physics.

TL;DR: The concept of lipid rafts as it has emerged from the study of synthetic membranes with the reality of lateral heterogeneity in biological membranes is compared.
Journal ArticleDOI

Coordination of Rho GTPase activities during cell protrusion

TL;DR: GTPase coordination in mouse embryonic fibroblasts is examined both through simultaneous visualization of two GTPase biosensors and using a ‘computational multiplexing’ approach capable of defining the relationships between multiple protein activities visualized in separate experiments, finding that RhoA is activated at the cell edge synchronous with edge advancement, whereas Cdc42 and Rac1 are activated 2 μm behind the edge with a delay of 40 s.
Journal ArticleDOI

Dynamic multiple-target tracing to probe spatiotemporal cartography of cell membranes.

TL;DR: An analytical single-particle tracking method and tool, multiple-target tracing (MTT), that takes advantage of the high spatial resolution provided by single-fluorophore sensitivity to generate dynamic maps at high densities of tracked particles, thereby providing global representation of molecular dynamics in cell membranes.
Journal ArticleDOI

Organization and Ca2+ Regulation of Adenylyl Cyclases in cAMP Microdomains

TL;DR: The regulation of many of the ACs by the ubiquitous second messenger Ca(2+) provides an overarching mechanism for integrating the activities of these two major signaling systems, and cAMP will exhibit distinct kinetics in discrete cellular domains.
References
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Journal ArticleDOI

Single-molecule imaging of the H-ras membrane-anchor reveals domains in the cytoplasmic leaflet of the cell membrane.

TL;DR: In this article, trajectories of individual molecules diffusing in the cytoplasmic leaflet of the plasma membrane were recorded, and the diffusion of individual membrane-anchored enhanced yellow fluorescent protein molecules was studied in live cells on timescales from 5 to 200 ms.
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Quantitative Coherent Anti-Stokes Raman Scattering Imaging of Lipid Distribution in Coexisting Domains

TL;DR: An image analysis model is developed and experimentally verified that can derive the relative molecular concentration from the difference of the two CARS intensities measured at the peak and dip frequencies of a CARS band.
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Lateral diffusion of GFP-tagged H2Ld molecules and of GFP-TAP1 reports on the assembly and retention of these molecules in the endoplasmic reticulum.

TL;DR: Lateral diffusion of GFP-tagged H2Ld molecules in the ER membrane reports on their interaction with the TAP complex during synthesis and peptide loading, suggesting that the T AP complex is very large, perhaps containing hundreds of proteins.
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Visualizing association of N-Ras in lipid microdomains: Influence of domain structure and interfacial adsorption

TL;DR: Two-photon fluorescence microscopy on giant unilamellar vesicles and tapping-mode atomic force microscopy are applied to follow the insertion of a fluorescently labeled and completely lipidated N-Ras protein into heterogeneous lipid bilayer systems, providing direct evidence that partitioning of N- Ras occurs preferentially into liquid-disordered lipid domains.
Journal ArticleDOI

Temporally resolved interactions between antigen-stimulated IgE receptors and Lyn kinase on living cells

TL;DR: In this article, the authors used fluorescence correlation spectroscopy (FCS) to investigate the mechanism of signaling-dependent protein-protein interactions in intact, living cells and reveal real-time interactions between Lyn and cross-linked FcepsilonRI implicated in downstream signaling events.
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